β-Hydroxybutyrate inhibits histone deacetylase 3 to promote claudin-5 generation and attenuate cardiac microvascular hyperpermeability in diabetes

Abstract

Aims/hypothesis Microvascular endothelial hyperpermeability, mainly caused by claudin-5 deficiency, is the initial pathological change that occurs in diabetes-associated cardiovascular disease. The ketone body β-hydroxybutyrate (BHB) exerts unique beneficial effects on the cardiovascular system, but the involvement of BHB in promoting the generation of claudin-5 to attenuate cardiac microvascular hyperpermeability in diabetes is poorly understood. Methods The effects of BHB on cardiac microvascular endothelial hyperpermeability and claudin-5 generation were evaluated in rats with streptozotocin-induced diabetes and in high glucose (HG)-stimulated human cardiac microvascular endothelial cells (HCMECs). To explore the underlying mechanisms, we also measured β-catenin nuclear translocation, binding of β-catenin, histone deacetylase (HDAC)1, HDAC3 and p300 to the Claudin-5 (also known as CLDN5) promoter, interaction between HDAC3 and β-catenin, and histone acetylation in the Claudin-5 promoter. Results We found that 10 weeks of BHB treatment promoted claudin-5 generation and antagonised cardiac microvascular endothelial hyperpermeability in rat models of diabetes. Meanwhile, BHB promoted claudin-5 generation and inhibited paracellular permeability in HG-stimulated HCMECs. Specifically, BHB (2 mmol/l) inhibited HG-induced HDAC3 from binding to the Claudin-5 promoter, although nuclear translocation or promoter binding of β-catenin did not change with BHB treatment. In addition, BHB prevented the binding and co-localisation of HDAC3 to β-catenin in HG-stimulated HCMECs. Furthermore, using mass spectrometry, acetylated H3K14 (H3K14ac) in the Claudin-5 promoter following BHB treatment was identified, regardless of whether cells were stimulated by HG or not. Although reduced levels of acetylated H3K9 in the Claudin-5 promoter were found following HG stimulation, increased H3K14ac was specifically associated with BHB treatment. Conclusions/interpretation BHB inhibited HDAC3 and caused acetylation of H3K14 in the Claudin-5 promoter, thereby promoting claudin-5 generation and antagonising diabetes-associated cardiac microvascular hyperpermeability.

Ketone body oxidation increases cardiac endothelial cell proliferation

“…Secondly, we addressed the possibility whether D‐β‐hydroxybutyrate would exert its functions by inhibiting class I histone deacetylases (HDAC) as this has been shown by treating mice with high doses (Shimazu et al , 2013 ). In ECs, there are contradictory reports about the efficiency of D‐β‐hydroxybutyrate acting as a HDAC inhibitor (Chriett et al , 2019 ; Li et al , 2021 ). To rule out the possibility that increased angiogenic potential of MCEC would merely be the consequence of HDAC inhibition, MCEC were treated with the well‐established HDAC inhibitor butyrate (Shimazu et al , 2013 ).…”

Exercise Ameliorates Atherosclerosis via Up-Regulating Serum β-Hydroxybutyrate Levels

“…It can bind to the G protein-coupled receptor GPR109A and activate the AMPK signal pathway to exert anti-inflammation effects and prevent the endoplasmic reticulum (ER) stress response and lipid accumulation [ 25 ]. BHB treatment can inhibit HDAC3 to promote claudin-5 generation and antagonize diabetes-associated cardiac microvascular hyperpermeability [ 50 ]. In addition, BHB can inhibit the activation of NLRP3 inflammasome, which is independent of GPR109A-mediated signaling, HDAC3, autophagy, or other starvation-regulated mechanisms [ 24 ].…”

Beta-Hydroxybutyrate: A Dual Function Molecular and Immunological Barrier Function Regulator

“…BHB was found as a competitive inhibiting catalytic site, directly inhibiting class I histone deacetylases (HDACs) ( 83 ), which were thought to participate in the regulation of gene expression by deacetylating lysine residues on histone and nonhistone proteins, such as NF-κB, TP53, MYC, and MYOD1 et al., and consequently regulates corresponding gene expression ( 84 , 85 ). Li and colleagues found that BHB upregulated claudin-5 gene expression and ameliorated the diabetes-associated cardiac endothelial hyperpermeability by inhibiting HDAC3 ( 60 ) ( Figure 2A ). By lysine β-hydroxybutyrylation, BHB can also directly modify proteins in multiple model organisms, including yeast, fly, rat, and human cells, and regulates gene expression ( 63 ).…”